LL Form Research Articles

Understanding impacts of a ransomware on medical and health facilities by utilizing LockBit as a case study

AbstractAll forms of data are meant to be secured. The emergence of a variety of techniques and tactics utilized by malware authors has made recovery difficult. LockBit ransomware has been in the news and the APT behind deployment of this ransomware virus has compromised several organizations. It is important to understand the behavior of such malware, to identify the anti‐analysis stuff it performs and to block it. The APT involved compromised several health and medical facilities including AIIMS in India and Sick Kids Hospital in Canada. Ransomwares are highly impactful family of malware. Impacts may begin from confidential medical information being harvested over a C2 and can go up to rendering medical equipment useless. This article discusses compromised entities and defending strategies that can be used to block LockBit ransomware using any generic SIEM tool. It also explains the need for tools that can survive ransomware encryption, a restart command‐line and can detect the behavioral patterns of such malware. Several samples were collected from various sources; the identities of compromise were collected from strings and behavior of those samples. Several detection mechanisms including YARA, SNORT, and generic HUNT rules have also been discussed.

Evaluation of the integrity of the facial muscles in leprosy patients using surface electromyography: a cross-sectional study

Objective This study sought to evaluate facial muscle integrity in clinical forms of leprosy through surface electromyography. Methods For evaluation of facial muscle integrity, 19 healthy subjects and 71 patients diagnosed with leprosy and under treatment in a National leprosy reference center were recruited. The muscles investigated, through surface electromyography, were the frontalis, orbicularis oculi, zygomaticus, masseter, and orbicularis oris. The evaluated features were Root Mean Square (RMS) and Mean frequency (Fmean). The facial muscles were analyzed jointly and separately for each leprosy clinical form and healthy individuals. Results Regarding the Fmean, there was a difference noted between the values of healthy subjects and patients of all clinical forms, except for the LL form (p = 0.986). The highest frequency values were seen in TT and BL forms, which did not present a significant difference between each other (p = 0.757). Asymmetry of the facial muscles was detected in TT (RMS) and TT, BB, and LL forms from the Fmean. RMS and Fmean indicated motor neurogenic impairment in facial muscles of all leprosy patients, except the LL form. Conclusions Surface electromyography is shown to be an auxiliary tool for the diagnosis of facial motor disorders imperceptible to clinical examination, helping to prevent impairments, which is fundamental for the social inclusion of leprosy patients.

Different cell death mechanisms are involved in leprosy pathogenesis

Leprosy is a chronic granulomatous disease that remains a serious public health problem in developing countries. According to the Madrid classification, leprosy presents in four clinical forms: two immunologically unstable forms (indeterminate and borderline) and two stable polar forms (tuberculoid and lepromatous). In leprosy, the relationship of cell death to clinical disease outcome remains unclear. Therefore, we investigated the extent of autophagy and different cell death mechanisms—such as apoptosis, necroptosis, and pyroptosis—in cutaneous lesions of patients with leprosy, as well as the role of these mechanisms in clinical disease progression. This cross-sectional analytical study included 30 patients with a confirmed diagnosis of leprosy, with 10 patients in each of the following groups: lepromatous (LL), tuberculoid (TT), and indeterminate (II) leprosy groups. For histopathological analysis, skin samples were subjected to haematoxylin-eosin staining and immunostaining for apoptotic and necroptotic markers. The results indicated that FasL expression was much higher in the LL form than in the TT and II forms. Similar results (higher expression in the LL form than in the TT and II forms) were observed for caspase 8, RIP1, and RIP3 expressions. MLKL, BAX, and caspase 3 expression levels were highest in the LL form, especially in globular foamy macrophages. Beclin-1 expression was highest in the TT form but was low in LL and II forms. Caspase 1 expression was highest in the LL form, followed by that in the TT and II forms. In conclusion, our study elucidates the role of different cell death mechanisms in the pathophysiology of various forms of leprosy and suggests measures that may be used to control the host response to infection and disease progression.

A critical realist perspective on humanness as a meaningful re‐entry of relational distinctions

AbstractAll forms of humanism that consider the human as a yardstick of society are marginalized by a Digital Technological Matrix that dominates the human, minimizes its role, and transforms it, to the point of dissolving its boundaries with the non‐human. Speaking of essence and human dignity still make sense? The thesis of this contribution is that the human is redefined and re‐evaluated precisely due to the process that makes the human ‘eccentric’ and places it in the system environment. The human re‐emerges as an intransitive essence endowed with an original, humble, relational dignity. The human can and must be regenerated through the relational reflexivity operated by relational subjects contextually capable of re‐entering within themselves the constitutive distinctions of their humanness, both individual and collective. The specific ‘intransitive’ essence of the human lies in the impossibility for it to renounce the meaning of its own existence. Precisely because of the confrontation with robotic technologies, the value of the human being and his dignity change.

Molecular epidemiology and transmission dynamics of leprosy among multicase families and case-contact pairs

BackgroundHuman-to-human transmission of Mycobacterium leprae among household contacts of active leprosy cases is significant, and surveillance of household contacts is vital to interrupting the transmission chain for this disease. This study was conducted to identify similarities in M. leprae strains, based on genomic single nucleotide polymorphisms (SNPs), among cases and their household contacts and in multicase families in order to decipher possible associations, transmission links, various clinical conditions of index cases that enhance person-to-person transmission, and timelines for transmission patterns. MethodsPCR for M. leprae DNA detection (amplification of the Rlep gene) and SNP subtyping of M. leprae strains was performed for 61 index cases and one of their household contacts. Additionally, we studied six families with multiple cases of leprosy, to understand timelines of infectivity and its relation to severity of the disease in the index cases. ResultsIndex cases with lepromatous (LL) and borderline lepromatous (BL) leprosy, together with a positive bacteriological index (BI) for M. leprae, result in a higher percentage of their contacts subclinically infected with M. leprae, with odds ratios (OR) of 6.6 (95% confidence interval (CI) 1.6–27.6) for BL and LL, and 7.07 (CI 1.41–35.41) for BI-positive index cases. 75% of the case–contact pairs had a similar SNP subtype of M. leprae. The timeline of infection in multicase families revealed that contacts were infected during the BI-positive period of the index case. ConclusionUsing molecular methods, we determined that positivity for M. leprae DNA in contacts of index leprosy cases was attributed to clinical characteristics of leprosy in the index cases. LL and BL forms of leprosy, together with positive BI, contributed to dissemination of infection to household contacts. In conclusion, we found a relationship between SNP subtypes within index case–contact pairs. This method can help decipher the transmission patterns and identify individuals at risk of contracting leprosy.

Electrochemistry for Life Detection on Ocean Worlds

AbstractAll forms of terrestrial life include cellular machinery to transform and regulate chemical energy flow through electron‐transfer pathways reliant upon key classes of biological redox molecules; life that evolved elsewhere, presumably, would possess a similar possibly overlapping set of energy‐management molecules. A second set of universal processes in terrestrial biology encompasses enzymatic change to add or remove functional groups, such as phosphate moieties, to biomolecules for a variety of purposes, from managing energy flow to tuning protein function; electrochemical assays can detect such enzymes. Thus, measurement of biological redox activity is a promising means to search for two major classes of life indicators as a component of future exploration missions to the “ocean worlds” of our solar system, those bodies that support substantial liquid oceans. Here, we assay a representative set of life‐critical redox molecules in synthetic seawater (SW) using electrochemical techniques. We also appropriate a well‐developed electrochemical assay that indicates the presence of phosphatase(s) by comparing the redox signatures of a substrate for, and the product of, the enzymatic process. We report measured limits of detection in SW as low as 10 nM for naturally occurring redox molecules and 3.1 aM for alkaline phosphatase over a 60‐minute period, demonstrating the promise and sensitivity of electrochemical sensors as effective life‐detection tools for future ocean worlds missions.

Beauty in the Neonatal Intensive Care Unit: Is Every Child a Pearl?

AbstractAll forms of beauty create appeal or enticement with moral significance. Sublime beauty draws one into a deep relationship that properly promotes the good and true. Parents tend to experience such beauty in their children, as eloquently described in works such as the 14th-century poem ‘The Pearl’, and they see this even when their children are desperately ill or dying. The experience of beauty in one’s child creates or reinforces the morality of caring. Unfortunately, at the end of modernity, the framing of beauty as only instrumental and subjective generally works against any recognition of dignity or respect for the very small pediatric patient. Practitioners who believe in the intrinsic value and dignity of persons as general concepts should recognize parental drawing toward their children as a particularization of a transcendental value.

Response of iNOS and its relationship with IL-22 and STAT3 in macrophage activity in the polar forms of leprosy

Leprosy is a chronic granulomatous infection that manifests as different clinical forms related to the immunological response. The aim of the study was to evaluated the response of IL-22, STAT3, CD68 and iNOS in leprosy skin lesions. The mean number IL-22 positive cells was 12.12±1.90cells/field in the TT form and 31.31±2.91cells/field in the LL form. STAT3 positive cells was 5.29±1.96 cells/field in the TT form, while this number was 11.13±3.48cells/field in the LL form. The mean number of CD68 positive cells was 25.18±6.21cells/field in the TT form and 62.81±8.13cells/field in the LL form. Quantitative analysis of iNOS revealed a significant difference, with the mean number of cells expressing the enzyme being 30.24±2.88cells/field in the TT form compared to 35.44±4.69cells/field in the LL form. Linear correlations in lesions of TT patients showed a moderate positive correlations between CD68 and iNOS, STAT3 and Inos, IL-22 and STAT3, and IL-22 and iNOS. Our results demonstrate that these factors can act synergistically to induce a microbicidal activity in the population of macrophages in the leprosy lesions.

Immunohistochemical analysis of the expression of cellular transcription NFκB (p65), AP-1 (c-Fos and c-Jun), and JAK/STAT in leprosy.

Leprosy is a disease whose clinical spectrum depends on the cytokine patterns produced during the early stages of the immune response. The main objective of this study was to describe the activation pattern of cellular transcription factors and to correlate these factors with the clinical forms of leprosy. Skin samples were obtained from 16 patients with the tuberculoid (TT) form and 14 with the lepromatous (LL) form. The histologic sections were immunostained with anti-c-Fos and anti-c-Jun monoclonal antibodies for investigation of AP-1, anti-NFκB p65 for the study of NFκB, and anti-JAK2, STAT1, STAT3, and STAT4 for investigation of the JAK/STAT pathway. Cells expressing STAT1 were more frequent in the TT form than in LL lesions (P = .0096), in agreement with the protective immunity provided by IFN-γ. STAT4 was also more highly expressed in the TT form than in the LL form (P = .0098). This transcription factor is essential for the development of a Th1 response because it is associated with interleukin-12. NFκB (p65) and STAT4 expression in the TT form showed a strong and significant correlation (r = 0.7556 and P = .0007). A moderate and significant correlation was observed between JAK2 and STAT4 in the TT form (r = 0.6637 and P = .0051), with these factors responding to interleukin-12 in Th1 profiles. The results suggest that STAT1, JAK2, and NFκB, together with STAT4, contribute to the development of cell-mediated immunity, which is able to contain the proliferation of Mycobacterium leprae.

Apoptotic activity and Treg cells in tissue lesions of patients with leprosy

In order to understand the apoptotic response and the participation of Treg cells in the spectral clinical evolution of leprosy, this study evaluated the immunohistochemical expression of caspase-3 and FoxP3 in skin lesions of leprosy patients with the polar forms of the disease. Forty-nine patients with a confirmed diagnosis of the disease were selected, including 27 with the TT form and 22 with the LL form. Quantitative analysis of caspase-3 immunostaining showed a higher expression of this mediator in the LL form (3.409 ± 0.6517 cells/mm2; p = 0.0001). Immunostaining for the transcription factor FoxP3 was higher in the LL form (3.891 ± 0.9294 cells/mm2; p = 0.0001). A moderate correlation between the two markers was observed in the TT form (r = 0.5214; p = 0.005). It can be concluded that Treg cells and apoptosis play an effective role for the host defense response, inducing mechanisms involved in the activation of cascades that interfere with the control of the immune response and cell homeostasis.

Abstract 2394: Impact of epidermal growth factor receptor (EGFR) gene intron 1 dinucleotide CA repeat polymorphism on protein expression and clinical outcome in oral cavity squamous cell carcinoma.

Abstract Epidermal growth factor receptor (EGFR) is one of the four members of the ErbB receptor family and has been shown to play a key role in tumorigenesis of many human cancers including oral cavity squamous cell carcinoma (OSCC). Previous studies have observed a significant relationship between EGFR overexpression and poor outcomes in head and neck squamous cell carcinoma (HNSCC) including OSCC. Elevated EGFR levels have primarily been attributed to enhanced transcriptional activity, rather than amplification of the gene. Since the EGFR intron 1 polymorphic CA dinucleotide repeats has been suggested to affect EGFR transcription efficiency, the present study was designed to explore the relationship between this CA repeats polymorphism, gene copy number and protein levels in Taiwanese OSCC. A total of 194 OSCCs were examined for EGFR protein overexpression using immunohistochemistry and for copy number using a fluorescence in situ hybridization (FISH) assay. DNAs from peripheral blood of 164 OSCC patients were also available for EGFR intron 1 CA dinucleotide repeats analysis. Overexpression and increased gene copy numbers of EGFR were found in 101 (52.06%) and 59 (30.41%) cases, respectively. The length of EGFR CA repeats were not associated with EGFR protein expression (p = 0.261) while the copy number was strongly related to protein overexpression (p < 0.001). EGFR protein overexpression was associated with young age (p = 0.036), tumor differentiation (p < 0.001) and lymph node metastasis (p = 0.028). EGFR gene copy number was also associated with young age (p = 0.009), tumor cell differentiation (p = 0.002), tumor depth (p = 0.001), lymph node metastasis (p = 0.051) and bone invasion (p = 0.003). However, the length of EGFR CA repeats was not associated with clinicopathologic factors. Patients with EGFR overexpression in their tumors were found to have significant poorer disease-free survival (DFS) (p = 0.026, log-rank test) and overall survival (OS) (p = 0.019, log-rank test) than those without overexpression. Patients with CA repeats SS form (both alleles < 19 repeats) had significantly shorter DFS and OS than those with SL and LL forms (both alleles ≥ 19 repeats) (p = 0.039 and 0.018 for DFS and OS, respectively). On the other hand, the association between an increase in EGFR gene copies and DFS or OS was insignificant. In summary, our current findings confirm that EGFR protein overexpression is closely related to EGFR copy number; but, do not support the role of CA repeats as an EGFR expression regulator. On the other hand, the length of CA repeats was significantly associated with both DFS and OS. Citation Format: Shiang-Fu Huang, I-How Chen, Chun-Ta Liao, Hung-Ming Wang, Ling-Ling Hsieh. Impact of epidermal growth factor receptor (EGFR) gene intron 1 dinucleotide CA repeat polymorphism on protein expression and clinical outcome in oral cavity squamous cell carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2394. doi:10.1158/1538-7445.AM2013-2394

Use of the polymerase chain reaction to detect Mycobacterium leprae in urine

Leprosy is an infectious disease caused by Mycobacterium leprae. The polymerase chain reaction (PCR) has been applied to detect M. leprae in different clinical samples and urine seems to be attractive for this purpose. PCR was used to improve the sensitivity for diagnosing leprosy by amplifying a 151-bp PCR fragment of the M. leprae pra gene (PCR-Pra) in urine samples. Seventy-three leprosy patients (39 males and 34 females, 14 to 78 years old) were selected for leprosy diagnosis at a reference laboratory in Maringá, PR, Brazil. Of these, 36 were under anti-leprosy multidrug therapy with dapsone and rifampicin for tuberculoid (TT) and dapsone, rifampicin and clofazimine for borderline (BB) and lepromatous (LL) forms. The control group contained 50 healthy individuals without any clinical history of leprosy. DNA isolated from leprosy patients' urine samples was successfully amplified by PCR-Pra in 46.6% (34/73) of the cases. The positivity of PCR-Pra for patients with the TT form was 75% for both patients under treatment and non-treated patients (P = 0.1306). In patients with the LL form, PCR-Pra positivity was 52 and 30% for patients under treatment and non-treated patients, respectively (P = 0.2386). PCR-Pra showed a statistically significant difference in detecting M. leprae between the TT and LL forms of leprosy in patients under treatment (P = 0.0033). Although the current study showed that the proposed PCR-Pra has some limitations in the detection of M. leprae, this method has the potential to be a useful tool for leprosy diagnosis mainly in TT leprosy where the AFB slit-skin smear is always negative.

Polymorphisms in the Serotonin Transporter Protein (SERT) Gene in Patients With Pulmonary Arterial Hypertension

Serotonin is a potent vasoconstrictor and pulmonary vascular growth factor whose concentration is increased in patients with pulmonary arterial hypertension (PAH). Its functions are mediated in part by the serotonin transporter protein (SERT) whose gene can have two allelic forms, both long (L) and short (S). The first was associated with greater function. ObjectivesTo determine whether the prevalence of the L allelic form of SERT is higher in patients with PAH than in the general population. To observe whether there are any clinical differences in patients with PAH based on the SERT allele. MethodsWe included patients diagnosed with PAH with catheterization based on the established criteria. Peripheral blood samples were taken and the DNA was extracted from the peripheral leukocytes. We amplified the promoter region of SERT by polymerase chain reaction and separated the products by electrophoresis. The patient samples were compared with samples from 50 healthy controls and among the most common types of PAH (idiopathic, thromboembolic and associated with connective tissue disorders). Several clinical variables were assessed according to the SERT gene alleles. ResultsThe study included 50 patients, and adequate samples were obtained in 49 patients (30 women). Mean age at diagnosis was 56±16 years. No differences were seen in the distribution of alleles between patients and controls (P=.54). There were no differences among the three most common types of PAH (P=.3). The most frequent allelic form was LS (54% patients, 56% controls). There were no differences in either age of diagnosis or response to treatment according to the SERT alleles. There was a trend toward higher mean pulmonary pressure levels in the LL forms (49±5mmHg vs 42±9mmHg, P=.07). ConclusionsThe distribution of SERT gene alleles does not appear to be different in patients with PAH than in the normal population. Different types of PAH have a similar distribution of alleles. The LL forms do not appear to confer either clinical differences or differences in response to treatment.

Polimorfismos en el gen de la proteína transportadora de serotonina (SERT) en pacientes con hipertensión arterial pulmonar

Serotonin is a potent vasoconstrictor and pulmonary vascular growth factor whose concentration is increased in patients with pulmonary arterial hypertension (PAH). Its functions are mediated in part by the serotonin transporter protein (SERT) whose gene can have two allelic forms, both long (L) and short (S). The first was associated with greater function. ObjectivesTo determine whether the prevalence of the L allelic form of SERT is higher in patients with PAH than in the general population. To observe whether there are any clinical differences in patients with PAH based on the SERT allele. MethodsWe included patients diagnosed with PAH with catheterization based on the established criteria. Peripheral blood samples were taken and the DNA was extracted from the peripheral leukocytes. We amplified the promoter region of SERT by polymerase chain reaction and separated the products by electrophoresis. The patient samples were compared with samples from 50 healthy controls and among the most common types of PAH (idiopathic, thromboembolic and associated with connective tissue disorders). Several clinical variables were assessed according to the SERT gene alleles. ResultsThe study included 50 patients, and adequate samples were obtained in 49 (30 women). Mean age at diagnosis was 56±16 years. No differences were seen in the distribution of alleles between patients and controls (P=.54). There were no differences among the three most common types of PAH (P=.3). The most frequent allelic form was LS (54% patients, 56% controls). There were no differences in either age of diagnosis or response to treatment according to the SERT alleles. There was a trend toward higher mean pulmonary pressure levels in the LL forms (49±5 vs 42±9mmHg, P=.07). ConclusionsThe distribution of SERT gene alleles does not appear to be different in patients with PAH than in the normal population. Different types of PAH have a similar distribution of alleles. The LL forms do not appear to confer either clinical differences or differences in response to treatment.

Immunoglobulin G and M Detection for Leprosy Diagnosis (129.18)

Abstract Leprosy is a curable disease that requires better diagnostic and prognostic tools to accompany preventive and therapeutic strategies. Circulating IgM antibodies against the M. leprae phenolic glycolipid-I (PGL-I) antigen have previously been used to assess humoral immunity. The presence of elevated titers of anti-PGL-I IgM reflects total bacterial load in the body; these antibodies, however, are generally low or absent in paucibacillary patients. Our objective was to compare 3 new recombinant antigens, ML0405, ML2331, and LID1 (comprising critical regions from ML0405 and ML2331), which recognize serum IgG, in comparison and in combination with PGL-I IgM. Serum samples were collected from 105 patients across the leprosy spectrum at the initial diagnosis and again after treatment. Patient positivity for LID1, ML0405, ML2331 and PGL-I tests was 67%, 62%, 65%, and 76%, respectively. A combination of the LID1 and PGL-1 antigens gave a positivity of 80%. After treatment, the ML0405 and PGL-I assays decreased their ELISA index values for all clinical forms, except for the LL form. Importantly, both antigens have been positively correlated with the bacillary load, clinical forms and the operational classification at diagnosis. Our results suggest that the combination of LID1 and PGL-I antigens, recognizing the IgG and IgM response, respectively, could be employed as an auxiliary tool for leprosy diagnosis. This work was funded in part by American Leprosy Mission.

The Functional State of the Complement System in Leprosy

Ninety-one patients with different clinical forms of leprosy, 36 lepromatous (LL), 33 tuberculoid (TL), and 22 dimorphic (DL), and 31 healthy volunteer donors were included in this study. Total complement system (CS) activity was assessed by hemolytic methods, whereas individual components were quantified by the enzyme-linked immunosorbent assay. Under conditions allowing initiation of cascade by the classic pathway (CP) but not alternative pathway (AP) activation, significant CS consumption was detected only in sera from patients with LL. In this group of patients, C4 but not factor B (fB) or C3 was significantly reduced, whereas mannose-binding lectin (MBL) serum levels were significantly higher. These results indicate that the CP is involved in CS activation in patients infected with Mycobacterium leprae manifesting LL clinical form of leprosy. An association is likely between circulating immune complexes and MBL high serum levels for initiation of CS activation in patients with LL form of leprosy.

Avaliação da concordância entre exames clínicos e laboratoriais no diagnóstico da hanseníase

This study examined the correlation between the clinical and laboratory diagnosis of leprosy, using biopsy results from laboratories 'A' and 'B' and the ML Flow test. Clinical and histopathological diagnoses presented 67.6% agreement. The laboratories showed 73.7% agreement in the bacterial index and laboratory 'B' detected 25.4% more positives. The highest agreement was in the LL form and lowest, in the I form. The highest diagnostic discrepancy was for the BB form. Clinical diagnosis agreement was 41.3% for laboratory 'A' and 54% for 'B'. The ML Flow test reclassified 10.7% of the patients. The spectrum of leprosy classification is important for a clearer understanding of the disease and its proper treatment, but is not used in health services, which use the simplified WHO criteria. This could be complemented by ML Flow testing. Such simplification is unacceptable for Leprosy Reference Centers regarding patient attendance, teaching and research, for which the standardization of the Ridley-Jopling classification is recommended.

Anti-leprosy protective vaccination of rhesus monkeys with BCG or BCG plus heat-killed Mycobacterium leprae: lepromin skin test results

Groups of rhesus monkeys (RM) were vaccinated and boosted with living Mycobacterium bovis Bacillus Calmette-Guerin (BCG) or BCG + low dose (LD) heat-killed Mycobacterium leprae (HKML) or high dose (HD) HKML or were unvaccinated. Animals vaccinated with BCG + LD and HD HKML were lepromin skin tested 2 weeks after boosting. All groups were lepromin tested 37 and 46 months after challenge with live M. leprae. Fernandez (72 h) and Mitsuda (28 day) responses were recorded. Ten of 10 rhesus monkeys in each of the two BCG + HKML-vaccinated groups significantly converted to strong positive Fernandez status within 2 weeks of boosting, compared to one of six positives in the unvaccinated unchallenged normal control group. Both BCG + HKML groups were significantly protected from clinical leprosy. Six of 10 in each of the two BCG + HKML groups significantly converted to Mitsuda positivity within 2 weeks of boosting compared to zero of six in the normal control group. The sizes of the Mitsuda responses were larger in the LD group than the HD HKML vaccinated/boosted group, suggesting suppression by vaccination with higher doses of HKML in combination with BCG. Fernandez responses were negative in normal RM as well as in the unvaccinated, ML-challenged group and the BCG-vaccinated, ML-challenged group at 37 or 46 months after ML inoculation, although the BCG-vaccinated group was significantly protected from leprosy and the unvaccinated group was not. In contrast, at 37 months the Fernandez reaction was positive in the BCG plus LD and the BCG plus HD HKML-vaccinated groups, both of which were significantly protected from clinical leprosy. By 46 months, the Fernandez responses were below significance in all groups. Thus, Fernandez reactivity is not a reliable correlate to protection from experimental leprosy in RM. Mitsuda responses became strongly positive in all four ML-challenged groups by 37 months and remained strongly positive at 46 months after ML inoculation, suggesting that strong Mitsuda reactivity reflects responses to living ML. BCG or BCG + LD or HD HKML vaccination/boosting of RM produced significant clinical protection from leprosy and there was a good correlation between protection from LL forms of leprosy and positive Mitsuda skin test responses after challenge with live ML. Positive Mitsuda responses were generated in essentially all individuals after challenge with live ML, and this response was primed by prior vaccination/boosting with BCG + HKML as shown by conversion to positivity 2 weeks after boosting. The data show that resistance to clinical leprosy is reflected by Mitsuda responses in ML-exposed RM, similar to results from human studies, and confirm the suitability of RM as a model for leprosy vaccine studies.

The MTHFR C677T, APOE, and PON55 gene polymorphisms show relevant interactions with cardiovascular risk factors.

Coronary artery disease (CAD) often cannot be explained by conventional risk factors (1). Various polymorphisms, such as MTHFR C677T, which causes hyperhomocysteinemia in the TT form (2), PON M55L, which causes increased paraoxonase (PON) activity in the LL form (3), and APOE , in which the presence of the e4 allele adversely affects lipoprotein metabolism (4), have been associated with CAD. Most research to date has focused on the individual effects of each polymorphism on the specific metabolic pathways in which they are known to participate. Discrepancies as to the importance of the effects of these polymorphisms on cardiovascular risk factors exist (5)(6)(7). The aim of this study was to investigate the combined effect of the presence of two or more of these polymorphisms on their corresponding cardiovascular risk factors. The study was performed in accordance with the guidelines of the institutions involved and was approved by the Hospital de Sant Joan and Jordi Gol Gorina Foundation ethics committees. All participants gave informed consent. Fasting blood samples were obtained from 400 volunteers between 0800 and 0900, following an overnight fast. Blood was collected (10 mL in each tube) in two EDTA-containing Vacutainer Tubes for plasma and leukocyte preparation and in an untreated tube for serum preparation. The tubes were kept at 4 °C until processing (within 2 h). Serum and plasma were stored in aliquots of 250 μL to 1 mL in cryotubes at −80 °C until required for analysis, and leukocytes were prepared from the remaining pellet, from which DNA was extracted. The genetic polymorphisms of the MTHFR , APOE , and PON 55 genes were analyzed according to previously described techniques (8)(9)(10)(11). Genotypes were identified independently by two observers, and the allele frequencies were calculated. The genotypes for …

Dependence of chiral recognition of α-amino acids and their derivatives on the nature of functional groups Calorimetric and spectroscopic studies

Calorimetric and nuclear relaxation time measurements were carried out at 25°C on binary and ternary aqueous solutions containing the L and D forms of the N-acetyl derivatives of the α-amino acids alanine, valine, leucine, methionine, aspartic acid and proline. The interaction coefficients of the virial expansion of the excess enthalpy were evaluated. Only the acetyl derivatives of leucine and proline present chiral recognition, namely a difference in the energetics of interaction between the LL and DL forms. This chiral recognition is larger in magnitude with respect to the corresponding free amino acids and changes sign. The role of the functional group is discussed and the results are interpreted using the preferential configuration model. Like–like interactions between functional groups seem to be responsible for the inversion of chiral recognition.